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Title: Structural Basis of Ubiquitin Recognition by the Ubiquitin-associated (UBA) Domain of the Ubiquitin Ligase EDD

Abstract

EDD (or HYD) is an E3 ubiquitin ligase in the family of HECT (homologous to E6-AP C terminus) ligases. EDD contains an N-terminal ubiquitin-associated (UBA) domain, which is present in a variety of proteins involved in ubiquitin-mediated processes. Here, we use isothermal titration calorimetry (ITC), NMR titrations, and pull-down assays to show that the EDD UBA domain binds ubiquitin. The 1.85{angstrom} crystal structure of the complex with ubiquitin reveals the structural basis of ubiquitin recognition by UBA helices {alpha}1 and {alpha}3. The structure shows a larger number of intermolecular hydrogen bonds than observed in previous UBA/ubiquitin complexes. Two of these involve ordered water molecules. The functional importance of residues at the UBA/ubiquitin interface was confirmed using site-directed mutagenesis. Surface plasmon resonance (SPR) measurements show that the EDD UBA domain does not have a strong preference for polyubiquitin chains over monoubiquitin. This suggests that EDD binds to monoubiquitinated proteins, which is consistent with its involvement in DNA damage repair pathways.

Authors:
; ; ; ; ;
Publication Date:
Research Org.:
Brookhaven National Laboratory (BNL) National Synchrotron Light Source
Sponsoring Org.:
DOE - OFFICE OF SCIENCE
OSTI Identifier:
1019677
Report Number(s):
BNL-95523-2011-JA
Journal ID: ISSN 0021-9258; JBCHA3; TRN: US201115%%317
DOE Contract Number:  
DE-AC02-98CH10886
Resource Type:
Journal Article
Resource Relation:
Journal Name: Journal of Biological Chemistry; Journal Volume: 282; Journal Issue: 49
Country of Publication:
United States
Language:
English
Subject:
08 HYDROGEN; BIOLOGICAL PATHWAYS; CALORIMETRY; CHAINS; CRYSTAL STRUCTURE; DNA DAMAGES; FUNCTIONALS; HYDROGEN; LIGASES; MUTAGENESIS; PLASMONS; PROTEINS; RESIDUES; RESONANCE; TITRATION; WATER; national synchrotron light source

Citation Formats

Kozlov, G., Nguyen, L, Lin, T, De Crescenzo, G, Park, M, and Gehring, K. Structural Basis of Ubiquitin Recognition by the Ubiquitin-associated (UBA) Domain of the Ubiquitin Ligase EDD. United States: N. p., 2007. Web. doi:10.1074/jbc.M705655200.
Kozlov, G., Nguyen, L, Lin, T, De Crescenzo, G, Park, M, & Gehring, K. Structural Basis of Ubiquitin Recognition by the Ubiquitin-associated (UBA) Domain of the Ubiquitin Ligase EDD. United States. doi:10.1074/jbc.M705655200.
Kozlov, G., Nguyen, L, Lin, T, De Crescenzo, G, Park, M, and Gehring, K. Mon . "Structural Basis of Ubiquitin Recognition by the Ubiquitin-associated (UBA) Domain of the Ubiquitin Ligase EDD". United States. doi:10.1074/jbc.M705655200.
@article{osti_1019677,
title = {Structural Basis of Ubiquitin Recognition by the Ubiquitin-associated (UBA) Domain of the Ubiquitin Ligase EDD},
author = {Kozlov, G. and Nguyen, L and Lin, T and De Crescenzo, G and Park, M and Gehring, K},
abstractNote = {EDD (or HYD) is an E3 ubiquitin ligase in the family of HECT (homologous to E6-AP C terminus) ligases. EDD contains an N-terminal ubiquitin-associated (UBA) domain, which is present in a variety of proteins involved in ubiquitin-mediated processes. Here, we use isothermal titration calorimetry (ITC), NMR titrations, and pull-down assays to show that the EDD UBA domain binds ubiquitin. The 1.85{angstrom} crystal structure of the complex with ubiquitin reveals the structural basis of ubiquitin recognition by UBA helices {alpha}1 and {alpha}3. The structure shows a larger number of intermolecular hydrogen bonds than observed in previous UBA/ubiquitin complexes. Two of these involve ordered water molecules. The functional importance of residues at the UBA/ubiquitin interface was confirmed using site-directed mutagenesis. Surface plasmon resonance (SPR) measurements show that the EDD UBA domain does not have a strong preference for polyubiquitin chains over monoubiquitin. This suggests that EDD binds to monoubiquitinated proteins, which is consistent with its involvement in DNA damage repair pathways.},
doi = {10.1074/jbc.M705655200},
journal = {Journal of Biological Chemistry},
number = 49,
volume = 282,
place = {United States},
year = {Mon Jan 01 00:00:00 EST 2007},
month = {Mon Jan 01 00:00:00 EST 2007}
}